US3374166A - Vertical gel electrophoresis apparatus - Google Patents

Vertical gel electrophoresis apparatus Download PDF

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Publication number
US3374166A
US3374166A US414654A US41465464A US3374166A US 3374166 A US3374166 A US 3374166A US 414654 A US414654 A US 414654A US 41465464 A US41465464 A US 41465464A US 3374166 A US3374166 A US 3374166A
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buffer solution
gel
tank
chamber
passageway
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US414654A
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Raymond Samuel
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor

Definitions

  • VERTICAL GEL ELECTROPHORESIS APPARATUS Filed Nov. so, 1964 United States ljatent 3,374,166 VERTICAL GEL ELECTROPHORESIS APPARATUS Samuel Raymond, 4312 @sage Ave., Philadelphia, Pa. 19104 Filed Nov. 30, 1964, Ser. No. 414,654 Claims priority, application Great Britain, Sept. 21, 1964, 38,461/64 7 IClaims. (Cl. 2041-299)
  • This invention relates to the art of electrophoresis and particularly .to improvements in apparatus for practicing vertical gel electrophoresis.
  • Electrophoresis generally, relates to the separation of a complex substance i-nto its component fractions by procedures based upon the migration or mobility of electrically charged fractions in a direct current electric field.
  • an electric eld developed between two spaced electrodes and a substance placed therein, the variously charged components or constituents of the substance move or migrate toward ythe respective electrodes of opposite charge and the respective components will move with different mobilities, i.e., at different rates.
  • separation may be accomplished on the various component fractions. This separation may take place on a suitable support medium such as acrylamide gel saturated with a buffer solution.
  • Each component separated by the electrophoresis procedure is subject to a qualitative and quantitative analysis and has, therefore, provided a useful tool in laboratory analysis of various substances such as albumin, enzymes, hemoglobin, carbohydrates, blood serum, etc., as well as various inorganic ions.
  • Vertical gel electrophoresis refers to electrophoresis wherein the component fractions migrate under the influence of an electric lield on a gel and in a vertical direction.
  • Vertical gel electrophoresis is generally known in the art as taught in U.S. Patent 3,129,158. This invention relates to improvements in the vertical gel apparatus of the known prior art.
  • This invention comprises attaching both electrodes to a single rigid component of the cell and providing a single .bipolar means for making electrical connection to both electrodes simultaneously.
  • Still another feature of this invention is the provision of means whereby samples may be introduced in-to the gel slab at a distance from the exposed end thereof.
  • FIG. l is an exploded perspective view of the vertical gel apparatus of this invention showing the two cornponent parts.
  • FIG. 2 is a sec-tional side elevational view .taken through the vertical gel apparatus assembled from the two component parts.
  • a vertical gel apparatus 1d includes an outer assembly 12- and an inner assembly 14, constructed of suitable electricaily insulating material such as a machinable plastic.
  • these two assemblies as shown in FIG. 1, are assembled in the manner illustrated in FIG. 2 they define a lower buffer solution containing tank 16,. an upper buffer solution containing tank 18.
  • Electrodes 20 and 22 are attached solely to inner member 14, and connecting insulated wires 21 and 23 are shown leading to a bipolar connector Z5' shown in FIG. l.
  • the two component parts define therebetween a vertical passageway 24 adapted to contain a gel support medium lfor the eleictrophoresis.
  • This ⁇ gel may be initially cast while the outer assembly 12 is in a horizontal position as disclosed in U.S. Patent 3,129,158.
  • the outer assembly 12 includes a back wall 26 with coolant passages 28 therein connected with suitable tubing connections 30 and 32.
  • a front wall 34 is provided with external tubing 36.
  • a bottom floor 37 supports the front and back walls as well as side walls 38 and 4t).
  • the side walls have a triangular shaped upper edge at 42 and 44 to add to the strength and rigidity of the assembly and maintain the back wall cooling plate in at shape.
  • Clamp straps 46 and 48 are provided at the midpoint of the outer component part 12.
  • a short top wall 45 is utilized to hold the gel during the casting thereof.
  • a slot 25 extends through the wall 26 between adjacent cooling channels 28. This slot allows a sample of a complex substance to be inserted into the gel between the ends thereof.
  • a close iitting plug 27 is provided to plug the slot 25 and is removable from the outside.
  • the inner assembly 14 includes a water cooled back wall having coolant passages S2 therein connected by suitable tubing 53, 55 to a source of coolant.
  • Side walls 54 and 56 are provided and they extend beyond the water cooled back wall lStb to provide a recessed area S8 extending vertically in the assembled component and providing the gel passageway 24.
  • the gel passageway is defined by the two side walls 54 and 56 as well as the back wall 50 of the inner assembly 14 and by the back wall 26 of the outer assembly 12. This arrangement prevents undue leakage of the gel being cast or gelled in passageway 24 as it eliminates butt joints at the edge of wall 50.
  • the side walls 54 and 56 are provided with triangular bottom edges 60 and 62 for rthe same purpose as edges 40 and 42.
  • Front wall 64 of lthe inner assembly 14 has external tubing 66 and a clamping strap 68 secured thereto so that this clamping strap will cooperate with clamping straps 46 and 48 and allow the assembly to be held together by a suitable clamp.
  • a floor '70 is liquid tight to hold the buffer solution in the upper tank 18.
  • plate Si) is provided with two spacers 49 and S1, one at each corner of said bottom edge.
  • a buffer drain tube 72 extends upwardly from the door 76 to a -distance equal to the desired height of the buffer solution in tank 18.
  • the tube 72 provides a path of uid communication through holes '73 in iioor 70 to the lower buffer solution containing tank 16.
  • External tubing 36 and 66 from the lower and upper buer solution containing tanks are connected to a pump 74 for the recirculation of buffer solution.
  • the recirculation is from the lower tank 16 to the upper tank 18 where it overows through the drain tube 16 and continually recirculates. This continuous recirculation during an electrophoresis run prevents alterations in buffer strength, composition or pH thereby maintaining more uniform conditions in electrophoresis cell and providing a more uniform and easily interpretable pattern than heretofore available.
  • the unit is first positioned with the back walls 26 and 50 horizontal and gel is poured into passage way 24. After gelling the assembly is turned upright to the position shown in FIG. 2 and the tanks 16 and 18 iilled with buffer solution. A substance to be separated is placed Iacross the gel at a suitable point. The electrodes and 22 are connected to a source of D C. and the pump 74 is started to recirculate Ithe buffer solution. Under the influence of the electric eld while on the permeable gel the components of the substance to be separated migrate at different rates and after a suitable period of time the electrophoresis may be stopped, the gel with the migrated separated components may be removed and then analyzed or interpreted.
  • a vertical gel electrophoresis apparatus of the type including a lower butter solution chamber capable of holding liquid butter solution, an upper buffer solution chamber capable of holding liquid buffer solution, a wall of each buffer solution chamber detining a gel passageway, the gel passageway being in fluid communication with liquid buffer solution in the buffer solution chambers and, when said passageway is lilled with impermeable gel free flow of buffer solution from the upper chamber to the lower chamber is prevented by said gel, an electrode in each butter solution chamber positioned below a normal level for butter solution therein, each electrode including means for connection to a source of D.C.
  • the upper and lower buffer solution chambers and the gel passage walls being constructed of a material having electrically insulating properties at least on the surface thereof, and means for holding the upper and lower buffer solution chambers in rigid assembled relationship, the improvements comprising, an internal drain tube extending from the bottom of the upper tank upwardly a distance equal to a desired level of buffer solution in the tank, external tubular connections establishing fluid communication Ibetween the tw-o tanks and pump means for recirculating bulier solution from the lower tank through the external tubular connections to the upper tank and then draining back to the lower tank.
  • a vertical gel electrophoresis apparatus as defined in claim 1 wherein the buiier solution carrying ⁇ tanks and gel containing passageway are provided by outer and inner assemblies and characterized by the electrodes for both buffer solution tanks being carried by one of the assemblies.
  • a vertical gel electrophoresis apparatus as defined in claim 2 in which the assembly carrying both electrodes is the inner assembly.
  • a vertical gel electrophoresis apparatus as dened in claim 2 and further characterized by a clamping strap at the center of the outer and inner component parts for clamping the outer and inner component parts in correctly assembled position.
  • a vertical gel electrophoresis apparatus as defined in claim 2, wherein walls of the two assemblies dening two walls of the gel containing passageway have cooling passages therein for the circulation of coolant and characterized by the inner of the two assemblies providing in addition the other two walls ofthe gel containing passageway.
  • a vertical gel electrophoresis apparatus as defined in claim 2 characterized by triangular shaped poitions side walls on the outer and inner assemblies.
  • a vertical gel electrophoresis cell as detined in claim 2 further characterized as having a slot means piercing an outer wall of said outer assembly between cooling channels thereof, said slot means being adaptable for the introduction of samples into the gel at a distance from an end thereof.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
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US414654A 1964-09-21 1964-11-30 Vertical gel electrophoresis apparatus Expired - Lifetime US3374166A (en)

Applications Claiming Priority (1)

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GB38461/64A GB1021287A (en) 1964-09-21 1964-09-21 Improvements in vertical gel electrophoresis apparatus

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3450624A (en) * 1966-07-20 1969-06-17 Fisher Scientific Co Apparatus for the separation of chemical components by the combination of electrophoresis and gel filtration
US3470080A (en) * 1966-06-27 1969-09-30 Samuel Raymond Multi-phase electrophoretic distribution
US3494846A (en) * 1968-06-11 1970-02-10 Pierre C Arquembourg Immuno-electrophoretic method and apparatus
US3719580A (en) * 1971-06-04 1973-03-06 R Roberts Electrophoretic apparatus
US3879280A (en) * 1974-04-16 1975-04-22 Us Health Gel slab electrophoresis cell and electrophoresis apparatus utilizing same
US3902986A (en) * 1972-04-29 1975-09-02 Colora Messtechnik Gmbh Apparatus for preparative electrophoresis
US3989612A (en) * 1974-12-26 1976-11-02 The Upjohn Company Elution device for gel electrophoresis
US4431506A (en) * 1982-09-07 1984-02-14 E-C Apparatus Corporation Apparatus for gel electrophoresis
US4588491A (en) * 1984-02-27 1986-05-13 International Biotechnologies, Inc. Horizontal gel electrophoresis device
US4612106A (en) * 1984-12-24 1986-09-16 Kromer Heiner M Cooling system for a slab gel electrophoresis apparatus
US4828669A (en) * 1988-04-28 1989-05-09 Eastman Kodak Company Electrophoresis device with removable buffer tank
US5427664A (en) * 1993-07-22 1995-06-27 Stoev; Stoyan V. Free solution electrophoresis-membrane filters trapping assay apparatus and method

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2677894B1 (fr) * 1991-06-20 1993-10-15 Bioprobe Systems Dispositif d'electrophorese.
GB2488838A (en) 2011-03-11 2012-09-12 Biomimetics Health Ind Ltd A stable antimicrobial aqueous hypochlorous acid solution

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3047489A (en) * 1961-05-25 1962-07-31 Raymond Samuel Apparatus for zone electrophoresis
US3129158A (en) * 1961-01-24 1964-04-14 Raymond Samuel Process for gel electrophoresis
US3208929A (en) * 1962-09-11 1965-09-28 Robert H Raymond Apparatus for gel electrophoresis
US3326790A (en) * 1962-01-04 1967-06-20 Messrs Lkb Produkter Ab Process and apparatus for vertical zone electrophoresis

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3129158A (en) * 1961-01-24 1964-04-14 Raymond Samuel Process for gel electrophoresis
US3047489A (en) * 1961-05-25 1962-07-31 Raymond Samuel Apparatus for zone electrophoresis
US3326790A (en) * 1962-01-04 1967-06-20 Messrs Lkb Produkter Ab Process and apparatus for vertical zone electrophoresis
US3208929A (en) * 1962-09-11 1965-09-28 Robert H Raymond Apparatus for gel electrophoresis

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3470080A (en) * 1966-06-27 1969-09-30 Samuel Raymond Multi-phase electrophoretic distribution
US3450624A (en) * 1966-07-20 1969-06-17 Fisher Scientific Co Apparatus for the separation of chemical components by the combination of electrophoresis and gel filtration
US3494846A (en) * 1968-06-11 1970-02-10 Pierre C Arquembourg Immuno-electrophoretic method and apparatus
US3719580A (en) * 1971-06-04 1973-03-06 R Roberts Electrophoretic apparatus
US3902986A (en) * 1972-04-29 1975-09-02 Colora Messtechnik Gmbh Apparatus for preparative electrophoresis
US3879280A (en) * 1974-04-16 1975-04-22 Us Health Gel slab electrophoresis cell and electrophoresis apparatus utilizing same
US3989612A (en) * 1974-12-26 1976-11-02 The Upjohn Company Elution device for gel electrophoresis
US4431506A (en) * 1982-09-07 1984-02-14 E-C Apparatus Corporation Apparatus for gel electrophoresis
US4588491A (en) * 1984-02-27 1986-05-13 International Biotechnologies, Inc. Horizontal gel electrophoresis device
US4612106A (en) * 1984-12-24 1986-09-16 Kromer Heiner M Cooling system for a slab gel electrophoresis apparatus
US4828669A (en) * 1988-04-28 1989-05-09 Eastman Kodak Company Electrophoresis device with removable buffer tank
US5427664A (en) * 1993-07-22 1995-06-27 Stoev; Stoyan V. Free solution electrophoresis-membrane filters trapping assay apparatus and method

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GB1021287A (en) 1966-03-02
DE1243647B (de) 1967-07-06

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